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目的研究浙江省近2年新分离的甲3型流行性感冒(流感)病毒株血凝素HA1基因特性,以了解浙江省流感病毒变异情况和WHO推荐疫苗株对人体的保护情况。方法采集浙江省近2年类流感病人的咽拭子和含漱液标本进行流感病毒的分离,对分离到的甲3型流感病毒进行核酸的提取,采用逆转录-聚合酶链反应(RT-PCR)扩增病毒基因后进行核苷酸序列测定,用DNA MAN和BioEdit生物软件对测序结果进行分析处理,并与WHO推荐疫苗株进行比对。结果近2年新分离到的甲3型流感病毒株HA1区域的核苷酸序列长度均为987bp,未发现核苷酸的丢失和插入,相应的点突变率为0.8%~1.3%,推导的氨基酸序列长度均为329个。2003年分离的毒株与同期WHO推荐使用的疫苗株A/Moscow/10/99比较,核苷酸同源性只有95.6%~95.9%,氨基酸总变异达20个,发生在抗原决定簇A、B区的有6个位点。2004年分离的毒株与同期WHO推荐使用的疫苗株A/fujian/411/2002(A/Ku-mamoto/102/2002,A Wyoming/3/2003)比较,核苷酸同源性达98.6%~99.1%,仅发生3~6个氨基酸的变异。基因进化树上,浙江省2003,2004年新分离的毒株均属于同一类性状的毒株,与2004年WHO推荐疫苗株接近,已远离2003年推荐疫苗株。结论浙江省甲3型流感病毒的抗原性已发生漂移,与我省流感的流行密切相关。2003年WHO推荐使用的甲3型流感疫苗对我省甲3型流感的预防效果不够理想,2004年推荐使用的疫苗对我省甲3型流感的预防效果较好。
Objective To study the HA1 gene characteristic of hemagglutinin (HA) gene of influenza A (H3N1) isolated in Zhejiang Province in recent two years so as to understand the variation of influenza virus in Zhejiang Province and the protection of the WHO vaccine against the human body. Methods Throat swabs and gargle samples of influenza-like patients in Zhejiang province were collected for the isolation of influenza virus. The nucleic acids were isolated from the influenza A virus type 3 virus by reverse transcription-polymerase chain reaction (RT- PCR) to amplify the nucleotide sequence of the virus gene. The sequencing results were analyzed by DNA MAN and BioEdit software and compared with the WHO recommended vaccine strains. Results The length of nucleotide sequence of influenza virus HA1 isolated in the recent 2 years was 987bp. No nucleotide loss or insertion was found, and the corresponding point mutation rate was 0.8% -1.3%. The deduced The amino acid sequence length is 329. Compared with the vaccine strain A / Moscow / 10/99 recommended by WHO in the same period in 2003, the nucleotide homology was only 95.6% -95.9% and the total amino acid variation was 20, which occurred in the antigenic determinants A, Area B has 6 sites. Compared with the vaccine strain A / fujian / 411/2002 (A / Ku-mamoto / 102/2002, A Wyoming / 3/2003) recommended by the WHO in the same period, nucleotide homology was 98.6% ~ 99.1%, only 3 to 6 amino acid mutations occurred. On the phylogenetic tree, the newly isolated strains in Zhejiang Province in 2003 and 2004 belonged to the same type of strains, which were close to the recommended vaccine strains in 2004 and far from the vaccine strains recommended in 2003. Conclusion The antigenicity of influenza A virus type 3 in Zhejiang Province has been drifting, which is closely related to the prevalence of influenza in our province. The type A influenza 3 vaccine recommended by the WHO in 2003 is not ideal for the prevention of influenza A type 3 influenza in our province. The vaccine recommended in 2004 has a good preventive effect on our influenza A type 3 influenza.